ω-3多不饱和脂肪酸预防多次七氟烷麻醉致新生小鼠脑神经毒性的机制:磷酸化Tau蛋白类淋巴系统清除途径  

Mechanism ofω-3 polyunsaturated fatty acids preventing sevoflurane-induced neurotoxicity in neonatal mice:phosphorylated Tau glymphatic system clearance pathway

作　　者：曹天宇 邸立超 黄培盈 曹珑璐 蒋素芳[1] 王雪吉 李佳琪 黄立宁[1] Cao Tianyu;Di Lichao;Huang Peiying;Cao Longlu;Jiang Sufang;Wang Xueji;Li Jiaqi;Huang Lining(Department of Anesthesiology,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Anesthesiology,Hebei General Hospital,Shijiazhuang 050051,China)

机构地区：[1]河北医科大学第二医院麻醉科,石家庄050000 [2]河北省人民医院麻醉科,石家庄050051

出　　处：《中华麻醉学杂志》2025年第2期170-177,共8页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(82071901);河北省自然科学基金精准医学联合重点项目(H2022206586);中央引导地方科技发展基金项目(246Z7725G)。

摘　　要：目的评价ω-3多不饱和脂肪酸预防多次七氟烷麻醉致新生小鼠脑神经毒性的机制与磷酸化Tau蛋白类淋巴系统清除途径的关系。方法 C57BL/6孕鼠18只, 采用随机数字表法分为2种喂养方式, 12只接受常规饲料喂养, 6只接受额外添加鱼油的饲料喂养[从妊娠第2天到分娩后14 d, 每20 g常规饲料添加ω-3多不饱和脂肪酸300 mg]。孕鼠分娩后, 取健康新生小鼠, 6日龄, 雌雄不拘, 体质量3~5 g。采用随机数字表法, 取6只常规饲料喂养孕鼠的新生小鼠48只纳入对照组(C组), 取6只常规饲料喂养孕鼠的新生小鼠48只纳入七氟烷麻醉组(S组), 取鱼油饲料喂养孕鼠的新生小鼠48只纳入ω-3多不饱和脂肪酸+七氟烷麻醉组(PS组)。子代小鼠出生后前21 d母乳喂养, 之后与母鼠分笼饲养, 自由进食常规饲料。S组和PS组子代小鼠于出生后6、7、8 d吸入3%七氟烷和40%氧气, 每天2 h。C组以相同的流速吸入40%氧气。于出生后33 d时行Y迷宫实验评估小鼠空间记忆和认知功能, 于出生后35 d时行转棒实验评估小鼠精细运动协调能力。于出生后14和35 d时, 采用小脑枕大池注射荧光示踪剂的方法, 评估类淋巴系统的流入功能(荧光示踪剂注射后30 min时渗透面积百分比)和流出功能(荧光示踪剂注射后90 min时残余面积百分比)。于出生后14 d, 采用Western blot法测定海马丝氨酸第202位点和苏氨酸第205位点磷酸化Tau蛋白(Tau-PS202/PT205)和总Tau蛋白表达;采集小鼠脑脊液标本, 采用ELISA法测定脑脊液磷酸化Tau蛋白浓度。于出生后35 d采用Western blot法测定海马caspase-3、caspase-9和细胞色素c(Cyt c)表达, 采用TUNEL法确定海马神经细胞凋亡率。结果与C组比较, S组新臂停留时间、新臂运动距离百分比和转棒停留时间减少, 出生后14 d时示踪剂渗透面积百分比减少, 示踪剂残余面积百分比增加, 海马Tau-PS202/PT205表达上调, 脑脊液磷酸化Tau蛋白浓�Objective To evaluate the relationship between the mechanism ofω-3 polyunsaturated fatty acids(ω-3 PUFAs)preventing sevoflurane-induced neurotoxicity and phosphorylated Tau glymphatic system clearance pathway in neonatal mice.MethodsEighteen C57BL/6 pregnant mice were used in this study and subjected to 2 feeding regiments using the random number table method.Twelve mice were selected to receive a standard diet,and 6 mice were selected to receive a diet supplemented with fish oil (ω-3 polyunsaturated fatty acids 300 mg was added to every 20 g of standard diet from the 2nd day of gestation to 14 days after parturition).The healthy neonatal mice of both sexes,aged 6 days,weighing 3-5 g,were selected after parturition.Forty-eight neonatal pups from 6 pregnant mice that were fed a standard diet were assigned to control group(C group),48 neonatal pups from 6 pregnant mice that were fed a standard diet were assigned to sevoflurane group(S group),and 48 neonatal pups from pregnant mice that were fed a diet supplemented with fish oil were assigned toω-3 PUFAs plus sevoflurane group(PS group)using the random number table method.All the offspring mice in all groups were breastfed until 21 days of birth and then were housed in separate cages from their mothers after 21 days of birth and provided with ad libitum access to standard food.S group and PS group inhaled 3%sevoflurane and 40%oxygen for 2 h daily on postnatal days 6,7 and 8.C group inhaled only 40%oxygen at the same flow rate.Y maze test was performed at postnatal day 33 to assess the spatial memory and cognitive function.The rotarod test was performed at postnatal day 35 to assess the fine motor coordination.The influx and efflux functions of the glymphatic system were assessed through intracisternal tracer infusion with the fluorescent tracer at postnatal days 14 and 35.The influx function was evaluated by the percentage of the area of tracer penetration 30 min after injection,while the efflux function was determined by the percentage of the residual area of t

关 键 词：脂肪酸类 ω3 七氟烷 神经胶细胞类淋巴系统 TAU蛋白质类 婴儿 新生 

分 类 号：R614[医药卫生—麻醉学]